DNMT3a Knockout CAR T Cells for Exhaustion Resistance
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Solution Overview
Problem
T cells engineered for adoptive cell therapy often undergo exhaustion, reducing their effectiveness against cancer and chronic infections due to epigenetic changes and immunosuppression by the tumor microenvironment, limiting their reinvigoration by checkpoint antibodies.
Innovation Solution
Generating T cells with a CRISPR-mediated knockout of the endogenous DNMT3A gene to downregulate gene expression, combined with exogenous T cell receptors or chimeric antigen receptors, which are resistant to exhaustion and can be reinvigorated by checkpoint antibodies.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Reliability
If T cells are engineered with exogenous TCR or CAR for adoptive cell therapy, then they can target cancer cells with desired specificity, but they undergo exhaustion and lose effector function over time
Solution Approach 1:
The patent extracts and removes the DNMT3A gene from the T cell genome using CRISPR/Cas9 technology. By taking out this specific gene responsible for de novo DNA methylation, the patent eliminates the mechanism that drives epigenetic exhaustion, thereby extending T cell persistence and maintaining therapeutic efficacy over time.
Solution Approach 2:
The patent changes the epigenetic parameter of DNA methylation by knocking out DNMT3A. This parameter change prevents the epigenetic reprogramming that leads to exhaustion, allowing T cells to maintain their effector function and central memory phenotype during long-term persistence in the tumor microenvironment.
2Reliability
If T cells are exposed to tumor microenvironment with PD-L1, then they can recognize and target cancer cells, but they become immunosuppressed and exhausted
Solution Approach 1:
The patent applies preliminary anti-action by preemptively knocking out DNMT3A before T cells encounter the tumor microenvironment. This pre-modification prevents the epigenetic changes that would normally occur upon exposure to PD-L1, thereby conferring resistance to immunosuppression and exhaustion before the harmful interaction takes place.
Solution Approach 2:
The patent converts the harmful effect of PD-L1 exposure into a benefit by using the CRISPR/Cas9 system to eliminate DNMT3A. The very exposure that would normally cause exhaustion is transformed into an opportunity to select and expand T cells with enhanced resistance properties, turning the harmful microenvironment into a beneficial selection pressure.
3Reliability
If checkpoint antibodies are administered to treat exhausted T cells, then they can block inhibitory receptors, but exhausted T cells cannot be fully reinvigorated due to epigenetic changes
Solution Approach 1:
The patent performs preliminary action by knocking out DNMT3A before administering checkpoint blockade therapy. This pre-modification of the epigenetic landscape prevents the establishment of exhaustion-associated methylation patterns, thereby enabling T cells to fully respond to and be reinvigorated by checkpoint antibodies that block PD-1 and other inhibitory receptors.
Data Source
AI summary
The present disclosure provides modified immune cells or precursors thereof (e.g., gene edited modified T cells) comprising an exogenous T cell receptor (TCR) and/or a chimeric antigen receptor (CAR) having specificity for a target antigen, and an insertion and/or deletion in an endogenous gene locus encoding DNMT3A. Compositions and methods of treatment are also provided.